USPTO Patents Application 10551504

Applicant
Serial No.
Filed
Page

Hiroyuki Tsunoda et al.
To Be Assigned
Herewith
3 of 13

Attorney's Docket No.: 14875-153US1 / C1-A0320Y2P-US

Amendments to the Claims :

This listing of claims replaces all prior versions and listings of claims in the application:
Listing of Claims :

1 . (original) An antibody comprising a single-chain polypeptide having binding activity
against TPO receptor (Mpl), wherein said antibody comprises two heavy chain variable regions
and two light chain variable regions.

2. (original) The antibody of claim 1, wherein the two heavy chain variable regions and
the two light chain variable regions are arranged in the order of heavy chain variable region, light
chain variable region, heavy chain variable region, and light chain variable region from the N
terminus of the single-chain polypeptide.

3. (currently amended) The antibody of claim 1 ef-2, wherein the two heavy chain
variable regions and the two light chain variable regions are linked by linkers.

4. (original) The antibody of claim 3, wherein the linkers comprise 15 amino acids.

5. (original) A chimeric antibody that binds to Mpl.

6. (original) The antibody of claim 5, which is a humanized antibody.

7. (currently amended) The antibody of claim 5 which is a minibody.

8. (original) An antibody that binds to soluble Mpl.

9. (original) An antibody that binds to human Mpl and monkey Mpl.

Applicant : Hiroyuki Tsunoda et al. Attorney's Docket No.: 14875-153US1 / C1-A0320Y2P-US

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10. (original) An antibody having agonistic activity against human Mpl and monkey

Mpl.

11. (original) An antibody whose binding activity to soluble Mpl is KD = 10" 6 M or

lower.

12. (original) An antibody whose binding activity to soluble Mpl is KD = 10" 7 M or

lower.

13. (original) An antibody whose TPO agonistic activity is EC50 = 100 nM or lower.

14. (original) An antibody whose TPO agonistic activity is EC50 = 30 nM or lower.

15. (original) An antibody whose TPO agonistic activity is EC50 = 10 nM or lower.

16. (original) An antibody which comprises a heavy chain variable region, wherein said
heavy chain variable regions comprises CDR1, CDR2 and CDR3 consisting of an amino acid
sequence of any one of:

[1] SEQ ID NOs: 3, 4, and 5
[2] SEQ ID NOs: 6, 7, and 8
[3] SEQ ID NOs: 9, 10, and 11
[4] SEQ ID NOs: 15, 16, and 17
[5] SEQ ID NOs: 18, 19, and 20
[6] SEQ ID NOs: 21, 22, and 23
[7] SEQ ID NOs: 24, 25, and 26
[8] SEQ ID NOs: 27, 28, and 29
[9] SEQ ID NOs: 30, 31, and 32

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[10] SEQ ID NOs: 33, 34, and 35
[11] SEQ ID NOs: 36, 37, and 38
[12] SEQ ID NOs: 39, 40, and 41
[13] SEQ ID NOs: 42, 43, and 44
[14] SEQ ID NOs: 48, 49, and 50
[15] SEQ ID NOs: 51, 52, and 53
[16] SEQ ID NOs: 54, 55, and 56
[17] SEQ ID NOs: 57, 58, and 59.

17. (original) An antibody which comprises a light chain variable region, wherein said
light chain variable region comprises CDR1, CDR2 and CDR3 consisting of an amino acid
sequence of any one of:

[1] SEQ ID NOs: 60, 61, and 62
[2] SEQ ID NOs: 63, 64, and 65
[3] SEQ ID NOs: 78, 79, and 80
[4] SEQ ID NOs: 84, 85, and 86
[5] SEQ ID NOs: 93, 94, and 95
[6] SEQ ID NOs: 96, 97, and 98
[7] SEQ ID NOs: 102, 103, and 104
[8] SEQ ID NOs: 108, 109, and 110
[9] SEQ ID NOs: 111, 112, and 113
[10] SEQ ID NOs: 114, 115, and 116.

18. (original) An antibody that comprises a heavy chain variable region and a light chain
variable region of any one of:

[1] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ID NOs: 3, 4, and 5, and a light chain variable

Applicant : Hiroyuki Tsunoda et al. Attorney's Docket No.: 14875-1 53US 1 / CI -A0320Y2P-US

Serial No. : To Be Assigned
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region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 60, 61, and 62;

[2] a heavy chain variable region that comprises CDR1 , CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ID NOs: 6, 7, and 8, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 63, 64, and 65;

[3] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ID NOs: 9, 10, and 11, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 63, 64, and 65;

[4] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 15, 16, and 17, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 63, 64, and 65;

[5] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 18, 19, and 20, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 63, 64, and 65;

[6] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 21, 22, and 23, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 78, 79, and 80;

[7] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 24, 25, and 26, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 63, 64, and 65;

[8] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 27, 28, and 29, and a light chain variable

Applicant: Hiroyuki Tsunoda et al. Attorney's Docket No.: 14875- 1 53US 1 / C1-A0320Y2P-US

Serial No. : To Be Assigned
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region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 84, 85, and 86;

[9] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 30, 31, and 32, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequence consisting
of SEQ ID NOs: 63, 64, and 65;

[10] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ID NOs: 33, 34, and 35, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 63, 64, and 65;

[1 1] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ID NOs: 36, 37, and 38, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 93, 94, and 95;

[12] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 39, 40, and 41, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 96, 97, and 98;

[13] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 42, 43, and 44, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 78, 79, and 80;

[14] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 45, 46, and 47, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 102, 103, and 104;

[15] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 48, 49, and 50, and a light chain variable

Applicant : Hiroyuki Tsunoda et al. Attorney's Docket No.: 14875- 1 53US1 / C1-A0320Y2P-US

Serial No. : To Be Assigned
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region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 63, 64, and 65;

[16] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 51, 52, and 53, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ID NOs: 1 08, 1 09, and 1 1 0,

[17] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ID NOs: 54, 55, and 56, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 comprising the amino acid sequences consisting
of SEQ ED NOs: 111, 112, and 113;

[18] a heavy chain variable region that comprises CDR1, CDR2, and CDR3 comprising
the amino acid sequences consisting of SEQ ED NOs: 57, 58, and 59, and a light chain variable
region that comprises CDR1, CDR2, and CDR3 each comprising the amino acid sequences
consisting of SEQ ED NOs: 1 14, 115, and 1 16.

19. (original) An antibody that comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ED NO: 118.

20. (original) An antibody that comprises a light chain variable region comprising the
amino acid sequence of SEQ ED NO: 120.

21 . (original) An antibody that comprises a heavy chain variable region comprising the
amino acid sequence of SEQ ED NO: 118 and a light chain variable region comprising the amino
acid sequence of SEQ ED NO: 120.

22. (original) An antibody comprising the amino acid sequence of SEQ ED NO: 122 or

264.

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Page

23. (original) An antibody that comprises a heavy chain variable region, wherein said
heavy chain variable region comprises FR1, FR2, FR3, and FR4 consisting of amino acid
sequences of any one of:

[1] SEQ ED NOs: 230, 232, 234, and 236
[2] SEQ ID NOs: 265, 267, 269, and 271
[3] SEQ ID NOs: 279, 281, 283, and 285
[4] SEQ ID NOs: 298, 299, 300, and 301
[5] SEQ ID NOs: 298, 299, 306, and 301.

24. (original) An antibody comprising a light chain variable region, wherein said light
chain variable region comprises FR1, FR2, FR3, and FR4 consisting of amino acid sequences of
any one of:

[1] SEQ ID NOs: 239, 241, 243, and 245
[2] SEQ ID NOs: 272, 274, 276, and 278
[3] SEQ ID NOs: 302, 303, 304, and 305
[4] SEQ ID NOs: 302, 307, 308, and 305.

25. (original) An antibody that comprises a heavy chain variable region and a light chain
variable region of any one of:

[1] a heavy chain variable region which comprises FR1, FR2, FR3, and FR4 having the
amino acid sequences consisting of SEQ ID NOs: 230, 232, 234, and 236, and a light chain
variable region which comprises FR1, FR2, FR3, and FR4 having the amino acid sequences
consisting of SEQ ID NOs: 239, 241, 243, and 245;

[2] a heavy chain variable region which comprises FR1, FR2, FR3, and FR4 having the
amino acid sequences consisting of SEQ ID NOs: 265, 267, 269, and 271, and a light chain
variable region which comprises FR1, FR2, FR3, and FR4 having the amino acid sequences
consisting of SEQ ID NOs: 272, 274, 276, and 278;

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Attorney's Docket No.: 14875-153US1 / C1-A0320Y2P-US

[3] a heavy chain variable region which comprises FR1, FR2, FR3, and FR4 having the
amino acid sequences consisting of SEQ ID NOs: 279, 281, 283, and 285, and a light chain
variable region which comprises FR1, FR2, FR3, and FR4 having the amino acid sequences
consisting of SEQ ID NOs: 272, 274, 276, and 278;

[4] a heavy chain variable region which comprises FR1, FR2, FR3, and FR4 having the
amino acid sequences consisting of SEQ ID NOs: 298, 299, 300, and 301, and a light chain
variable region which comprises FR1, FR2, FR3, and FR4 having the amino acid sequences
consisting of SEQ ID NOs: 302, 303, 304, and 305;

[5] a heavy chain variable region which comprises FR1, FR2, FR3, and FR4 having the
amino acid sequences consisting of SEQ ID NOs: 298, 299, 306, and 301, and a light chain
variable region which comprises FR1, FR2, FR3, and FR4 having the amino acid sequences
consisting of SEQ ID NOs: 302, 307, 308, and 305.

26. (original) An antibody that comprises a heavy chain variable region, wherein said
heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 229, 256, 262,
289, or 295.

27. (original) An antibody that comprises a light chain variable region, wherein said
light chain variable region comprises the amino acid sequence of SEQ ED NO: 238, 258, 291, or
297.

28. (original) An antibody that comprises a heavy chain variable region and a light chain
variable region of any one of:

[1] a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:
229, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 238;

[2] a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:
256, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 258;

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Attorney's Docket No.: 14875-153US1 / C1-A0320Y2P-US

[3] a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:
262, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 258;

[4] a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:
289, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 291 ;

[5] a heavy chain variable region comprising the amino acid sequence of SEQ ED NO:
295, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 297.

29. (original) An antibody that comprises the amino acid sequence of SEQ ED NO: 2,
254, 260, 287, or 293.

30. (currently amended) An antibody having an activity equivalent to that of an antibody
of any on e of claims claim 16 to 29 , wherein said antibody comprises the amino acid sequence
set forth in any on e of claims claim 1 6 to 29 , in which one or more amino acids have been
substituted, deleted, added and/or inserted.

3 1 . (currently amended) An antibody that recognizes an epitope recognized by an
antibody of any on e of claims claim 16 to 30 .

32. (original) An antibody that recognizes the region of amino acids 26 to 274 of human

Mpl.

33. (currently amended) An antibody of any on e of claims claim l-te-32, which has TPO
agonistic activity.

34. (currently amended) A polynucleotide encoding an antibody of any on e of claims
claim 1 to 33 .

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Attorney's Docket No.: 14875-153US1 / C1-A0320Y2P-US

35. (currently amended) A polynucleotide hybridizing to the polynucleotide of claim 34
under stringent conditions, wherein said polynucleotide encodes an antibody having binding
activity against TPO receptor (Mpl) activity e quival e nt to that of an antibody of any on e of
claims 1 to 33 .

36. (currently amended) A vector comprising the polynucleotide of claim 34 or 35 .

37. (currently amended) A host cell that carries the polynucleotide of claim 34 or 35, or
th e v e ctor of claim 36 .

38. (currently amended) A pharmaceutical composition comprising an antibody of any
on e of claims claim 1 to 33 .